一种简单的人脐带间充质干细胞分离培养方法

A simple method for isolation of human umbilical cord mesenchymal stem cell

目的建立一种简单的人脐带间充质干细胞分离培养方法。方法取新鲜脐带,剪成5cm长的小段,直接剪碎为糊状,加入含10﹪胎牛血清的DMEM/F12在培养瓶中培养,光学显微镜下观察细胞的生长特征,运用流式细胞仪检测分析细胞的抗原标志表达,并检测其体外多向分化潜能。结果运用不剥离血管组织、不用酶消化的组织贴块培养法可以从新鲜脐带中分离到贴壁生长的成纤维细胞样细胞,这些细胞表达CD29、CD44、CD90和CD105,并可在体外诱导分化为脂肪、骨和软骨细胞。结论运用不剥离血管组织、不用酶消化的组织贴块培养法可以从人脐带中分离到间充质干细胞,其生物学特性与骨髓间充质干细胞相似。

Objective To establish a simple method for isolation of human umbilical cord mesenchymal stem cell. Methods Five-em long segments from fresh umbilical cord were directly minced into pasty fine pieces, and the pieces were cultured in tissue flask with DMEM/F 12 medium supplemented with 10 % fetal bovine serum. The morphological characteristic of the cells was investigated by light microscope, and their expression of mesenchymal surface markers were analyzed by flow cytometry. The multidifferentiation potential was examined in vitro, too. Results The fibroblast-like cells could be isolated from the fresh umbilical cord by a nonenzymatie isolation procedure without removing of the vessels. These adherent passage 3 cells expressed mesenchymal markers including CD29, CD44, CD90 and CD105, and also could be induced to differentiate into adipocytes, osteoblasts and chondrocytes. Conclussion Mesenchymal stem cells, which biological characteristic was similar to bone marrow mesenchymal stem cells, could be isolated from fresh human umbilical cord by using a nonenzymatic isolation procedure without removing of the vessels.